Difference between revisions of "Part:BBa K934022"
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This part produces LasI enzyme in the presence of LuxR-3OC6HSL complex. | This part produces LasI enzyme in the presence of LuxR-3OC6HSL complex. | ||
− | [[Image:Plux-LasI_result.png|thumb|center|500px| | + | [[Image:Plux-LasI_result.png|thumb|center|500px|Fig. 1]] |
To characterize Plux-LasI (BBa_K934022), we introduced Plux-LasI pSB3K3 with Ptet-LuxR ([https://parts.igem.org/Part:BBa_S03119 BBa_S03119]) on pSB6A1 to ''E.coli'' as “3OC6HSL dependent 3OC12HSL producer cell”. In this ''E.coli'', constitutively expressed LuxR activates the expression of LasI in the presence of 3OC6HSL. We then introduced Ptrc-LasR and Plas-GFP ([https://parts.igem.org/Part:BBa_K649001 BBa_K649001]) to ''E.coli'' as a “Las reporter cell”. | To characterize Plux-LasI (BBa_K934022), we introduced Plux-LasI pSB3K3 with Ptet-LuxR ([https://parts.igem.org/Part:BBa_S03119 BBa_S03119]) on pSB6A1 to ''E.coli'' as “3OC6HSL dependent 3OC12HSL producer cell”. In this ''E.coli'', constitutively expressed LuxR activates the expression of LasI in the presence of 3OC6HSL. We then introduced Ptrc-LasR and Plas-GFP ([https://parts.igem.org/Part:BBa_K649001 BBa_K649001]) to ''E.coli'' as a “Las reporter cell”. | ||
− | + | The supernatants of the cultures of Plux-LasI cell were used as the inducer in the reporter assay. | |
'''We accomplished a positive feedback system with our improved part Plux-LasI([https://parts.igem.org/Part:BBa_K934012 BBa_K934012]).''' | '''We accomplished a positive feedback system with our improved part Plux-LasI([https://parts.igem.org/Part:BBa_K934012 BBa_K934012]).''' |
Revision as of 08:49, 28 October 2012
Plux-LasI
We constructed this part by combining BBa_R0062 and BBa_K081016. This part produces LasI enzyme in the presence of LuxR-3OC6HSL complex.
To characterize Plux-LasI (BBa_K934022), we introduced Plux-LasI pSB3K3 with Ptet-LuxR (BBa_S03119) on pSB6A1 to E.coli as “3OC6HSL dependent 3OC12HSL producer cell”. In this E.coli, constitutively expressed LuxR activates the expression of LasI in the presence of 3OC6HSL. We then introduced Ptrc-LasR and Plas-GFP (BBa_K649001) to E.coli as a “Las reporter cell”.
The supernatants of the cultures of Plux-LasI cell were used as the inducer in the reporter assay.
We accomplished a positive feedback system with our improved part Plux-LasI(BBa_K934012).
For more information, see [http://2012.igem.org/Team:Tokyo_Tech/Project our work in Tokyo_Tech 2012 wiki].
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 745
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 307
- 1000COMPATIBLE WITH RFC[1000]